Cyclohexanone monooxygenase (CHMO) was used as a model enzyme to find suitable freeze-drying conditions for long-term storage of an isolated monooxygenase. CHMO is a Baeyer-Villiger monooxygenase (BVMO) known for its ability to catalyze a large number of oxidation reactions. With a focus on establishing the optimal formulation, additives were tested for enzyme stabilization during and after lyophilization. The results were successfully transferred to two other monooxygenases, namely the BVMO cyclopentadecanone monooxygenase (CPDMO) and a cytochrome P450 monooxygenase, P450 BM3. In the absence of a lyoprotectant, lyophilized P450 BM3 is almost completely inactivated, while the lyophilized BVMOs quickly lost activity when stored at 50°C. Lyophilization in the presence of 2% (w/v) sucrose was found to be the best formulation to preserve activity and protect against inactivation when stored as lyophilizate at 50°C.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.jbiotec.2015.03.010 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Unlocking the potential of cyanobacteria: a high-throughput strategy for enhancing biocatalytic performance through genetic optimization.
Trends Biotechnol
December 2024
Institute of Applied Synthetic Chemistry, TU Wien, Getreidemarkt 9, 1060, Vienna, Austria. Electronic address:
Cyanobacteria show promise as hosts for whole-cell biocatalysis. Their photoautotrophic metabolism can be leveraged for a sustainable production process. Despite advancements, performance still lags behind heterotrophic hosts.
View Article and Find Full Text PDFStructure-Guided Evolution of Cyclohexanone Monooxygenase Toward Bulky Omeprazole Sulfide: Substrate Migration and Stereoselectivity Inversion.
Chemphyschem
June 2024
Department: Key laboratory of industrial Biotechnology. Institution: Ministry of Education, School of Biotechnology, Address, Jiangnan University, Wuxi, 214122, Jiangsu, China.
Structure-guided engineering of a CHMO from Amycolatopsis methanolica (AmCHMO) was performed for asymmetric sulfoxidation activity and stereoselectivity toward omeprazole sulfide. Initially, combinatorial active-site saturation test (CASTing) and iteratively saturation mutagenesis (ISM) were performed on 5 residues at the "bottleneck" of substrate tunnel, and MT3 was successfully obtained with a specific activity of 46.19 U/g and R-stereoselectivity of 99 % toward OPS.
View Article and Find Full Text PDFIntermediate product control in cascade reaction for one-pot production of ε-caprolactone by Escherichia coli.
Biotechnol J
February 2024
School of Biology and Biological Engineering, South China University of Technology, Guangzhou, China.
ε-Caprolactone is an important non-toxic compound for polymer synthesis like polycaprolactone which has been widely used in drug delivery and degradable plastics. To meet the demand for a green economy, a bi-enzymatic cascade, consisting of an alcohol dehydrogenase (ADH) and a cyclohexanone monooxygenase (CHMO), was designed and introduced into Escherichia coli to synthesize ε-caprolactone from cyclohexanol with a self-sufficient NADPH-cofactor regeneration system. To further improve the catalytic efficiency, a carbonyl group-dependent colorimetric method using inexpensive 2,4-dinitrophenylhydrazine (DNPH) was developed for assay of cyclohexanone, an intermediate production of cascade reaction.
View Article and Find Full Text PDFImproving the Enantioselectivity of CHMO for Asymmetric Synthesis of Podophyllotoxin Precursor.
Chembiochem
December 2023
State Key Laboratory of Bioreactor Engineering, Shanghai Collaborative Innovation Center for Biomanufacturing, School of Biotechnology, East China University of Science and Technology, 130 Meilong Road, Shanghai, 200237, China.
(R)-β-piperonyl-γ-butyrolactones are key building blocks for the synthesis of podophyllotoxin, which have demonstrated remarkable potential in cancer treatment. Baeyer-Villiger monooxygenases (BVMOs)-mediated asymmetric oxidation is a green approach to produce chiral lactones. While several BVMOs were able to oxidize the corresponding cyclobutanone, most BVMOs gave the (S) enantiomer while Cyclohexanone monooxygenase (CHMO) from Brevibacterium sp.
View Article and Find Full Text PDFPhotoinduced Promiscuity of Cyclohexanone Monooxygenase for the Enantioselective Synthesis of α-Fluoroketones.
Angew Chem Int Ed Engl
December 2022
Center of Chemistry for Frontier Technologies, Department of Chemistry, Zhejiang University, Hangzhou, 310027, China.
The development of mild, efficient, and enantioselective methods for preparing chiral fluorinated compounds has been a long-standing challenge. Herein, we report a promiscuous cyclohexanone monooxygenase (CHMO) for the photoinduced synthesis of chiral α-fluoroketones via enantioselective reductive dehalogenation of α,α-halofluoroketones. Wild-type CHMO from Acinetobacter sp.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!